CN103467729B - Polyethylene glycol-amino acid-polyester three-block polymkeric substance and its production and use - Google Patents
Polyethylene glycol-amino acid-polyester three-block polymkeric substance and its production and use Download PDFInfo
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- CN103467729B CN103467729B CN201310345062.1A CN201310345062A CN103467729B CN 103467729 B CN103467729 B CN 103467729B CN 201310345062 A CN201310345062 A CN 201310345062A CN 103467729 B CN103467729 B CN 103467729B
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- polyethylene glycol
- triblock polymer
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- polyester
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- -1 Polyethylene Polymers 0.000 title claims abstract description 136
- 239000004698 Polyethylene Substances 0.000 title claims abstract description 135
- 229920000573 polyethylene Polymers 0.000 title claims abstract description 135
- 229920000728 polyester Polymers 0.000 title claims abstract description 52
- 239000000126 substance Substances 0.000 title claims abstract description 9
- 229920000642 polymer Polymers 0.000 claims abstract description 82
- 239000000693 micelle Substances 0.000 claims abstract description 65
- 239000002245 particle Substances 0.000 claims abstract description 40
- 239000003814 drug Substances 0.000 claims abstract description 20
- 239000002114 nanocomposite Substances 0.000 claims abstract description 16
- 238000011161 development Methods 0.000 claims abstract description 6
- 230000004048 modification Effects 0.000 claims abstract description 4
- 238000012986 modification Methods 0.000 claims abstract description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 96
- 238000006243 chemical reaction Methods 0.000 claims description 94
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 78
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 68
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims description 52
- 238000002360 preparation method Methods 0.000 claims description 50
- 239000002159 nanocrystal Substances 0.000 claims description 49
- 239000000243 solution Substances 0.000 claims description 47
- 229960001701 chloroform Drugs 0.000 claims description 41
- 239000002904 solvent Substances 0.000 claims description 36
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 33
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 30
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 30
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 28
- 239000002244 precipitate Substances 0.000 claims description 27
- 229910052786 argon Inorganic materials 0.000 claims description 26
- 239000008346 aqueous phase Substances 0.000 claims description 22
- 238000002390 rotary evaporation Methods 0.000 claims description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 20
- 239000000047 product Substances 0.000 claims description 19
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 18
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 17
- 229940079593 drug Drugs 0.000 claims description 16
- 239000003960 organic solvent Substances 0.000 claims description 16
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 15
- UKVIEHSSVKSQBA-UHFFFAOYSA-N methane;palladium Chemical compound C.[Pd] UKVIEHSSVKSQBA-UHFFFAOYSA-N 0.000 claims description 15
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 14
- 230000002209 hydrophobic effect Effects 0.000 claims description 14
- SZVJSHCCFOBDDC-UHFFFAOYSA-N iron(II,III) oxide Inorganic materials O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 claims description 14
- 229910052757 nitrogen Inorganic materials 0.000 claims description 14
- 229920001223 polyethylene glycol Polymers 0.000 claims description 14
- 239000002253 acid Substances 0.000 claims description 13
- 238000005303 weighing Methods 0.000 claims description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 12
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 12
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 12
- 239000002202 Polyethylene glycol Substances 0.000 claims description 11
- 239000000203 mixture Substances 0.000 claims description 11
- 239000012074 organic phase Substances 0.000 claims description 11
- UHYPYGJEEGLRJD-UHFFFAOYSA-N cadmium(2+);selenium(2-) Chemical compound [Se-2].[Cd+2] UHYPYGJEEGLRJD-UHFFFAOYSA-N 0.000 claims description 10
- 239000001257 hydrogen Substances 0.000 claims description 10
- 229910052739 hydrogen Inorganic materials 0.000 claims description 10
- 239000011261 inert gas Substances 0.000 claims description 10
- 239000000178 monomer Substances 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 9
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 claims description 8
- 238000007385 chemical modification Methods 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 8
- 230000001376 precipitating effect Effects 0.000 claims description 8
- KSBAEPSJVUENNK-UHFFFAOYSA-L tin(ii) 2-ethylhexanoate Chemical compound [Sn+2].CCCCC(CC)C([O-])=O.CCCCC(CC)C([O-])=O KSBAEPSJVUENNK-UHFFFAOYSA-L 0.000 claims description 8
- VGALFAWDSNRXJK-VIFPVBQESA-N L-aspartic acid beta-benzyl ester Chemical compound OC(=O)[C@@H](N)CC(=O)OCC1=CC=CC=C1 VGALFAWDSNRXJK-VIFPVBQESA-N 0.000 claims description 7
- 238000004132 cross linking Methods 0.000 claims description 7
- PAPBSGBWRJIAAV-UHFFFAOYSA-N ε-Caprolactone Chemical group O=C1CCCCCO1 PAPBSGBWRJIAAV-UHFFFAOYSA-N 0.000 claims description 7
- 125000003172 aldehyde group Chemical group 0.000 claims description 6
- 125000000304 alkynyl group Chemical group 0.000 claims description 6
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 6
- 229940024606 amino acid Drugs 0.000 claims description 6
- 150000001413 amino acids Chemical class 0.000 claims description 6
- 150000001540 azides Chemical class 0.000 claims description 6
- 239000007853 buffer solution Substances 0.000 claims description 6
- 238000012650 click reaction Methods 0.000 claims description 6
- 238000006482 condensation reaction Methods 0.000 claims description 6
- 238000005886 esterification reaction Methods 0.000 claims description 6
- 150000004820 halides Chemical class 0.000 claims description 6
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 6
- 239000012071 phase Substances 0.000 claims description 6
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 6
- KIDHWZJUCRJVML-UHFFFAOYSA-N putrescine Chemical compound NCCCCN KIDHWZJUCRJVML-UHFFFAOYSA-N 0.000 claims description 6
- BGGHCRNCRWQABU-JTQLQIEISA-N (2s)-2-amino-5-oxo-5-phenylmethoxypentanoic acid Chemical compound OC(=O)[C@@H](N)CCC(=O)OCC1=CC=CC=C1 BGGHCRNCRWQABU-JTQLQIEISA-N 0.000 claims description 5
- 239000003153 chemical reaction reagent Substances 0.000 claims description 5
- QBPPRVHXOZRESW-UHFFFAOYSA-N 1,4,7,10-tetraazacyclododecane Chemical compound C1CNCCNCCNCCN1 QBPPRVHXOZRESW-UHFFFAOYSA-N 0.000 claims description 4
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 claims description 4
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 4
- 229960004424 carbon dioxide Drugs 0.000 claims description 4
- 235000011089 carbon dioxide Nutrition 0.000 claims description 4
- 239000003431 cross linking reagent Substances 0.000 claims description 4
- 238000001704 evaporation Methods 0.000 claims description 4
- 229910052734 helium Inorganic materials 0.000 claims description 4
- 239000001307 helium Substances 0.000 claims description 4
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 claims description 4
- 239000003446 ligand Substances 0.000 claims description 4
- 229910021645 metal ion Inorganic materials 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- 150000003384 small molecules Chemical class 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- 238000001291 vacuum drying Methods 0.000 claims description 4
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 3
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 3
- IYMAXBFPHPZYIK-BQBZGAKWSA-N Arg-Gly-Asp Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IYMAXBFPHPZYIK-BQBZGAKWSA-N 0.000 claims description 3
- KAFHLONDOVSENM-HNNXBMFYSA-N O-Benzyl-L-tyrosine Chemical compound C1=CC(C[C@H](N)C(O)=O)=CC=C1OCC1=CC=CC=C1 KAFHLONDOVSENM-HNNXBMFYSA-N 0.000 claims description 3
- 229940009456 adriamycin Drugs 0.000 claims description 3
- MHDVGSVTJDSBDK-UHFFFAOYSA-N dibenzyl ether Chemical compound C=1C=CC=CC=1COCC1=CC=CC=C1 MHDVGSVTJDSBDK-UHFFFAOYSA-N 0.000 claims description 3
- 239000007850 fluorescent dye Substances 0.000 claims description 3
- JJTUDXZGHPGLLC-UHFFFAOYSA-N lactide Chemical compound CC1OC(=O)C(C)OC1=O JJTUDXZGHPGLLC-UHFFFAOYSA-N 0.000 claims description 3
- 230000008685 targeting Effects 0.000 claims description 3
- NKIJBSVPDYIEAT-UHFFFAOYSA-N 1,4,7,10-tetrazacyclododec-10-ene Chemical compound C1CNCCN=CCNCCN1 NKIJBSVPDYIEAT-UHFFFAOYSA-N 0.000 claims description 2
- 102000020313 Cell-Penetrating Peptides Human genes 0.000 claims description 2
- 108010051109 Cell-Penetrating Peptides Proteins 0.000 claims description 2
- 229910002518 CoFe2O4 Inorganic materials 0.000 claims description 2
- 229910005335 FePt Inorganic materials 0.000 claims description 2
- 229910052688 Gadolinium Inorganic materials 0.000 claims description 2
- 229910017163 MnFe2O4 Inorganic materials 0.000 claims description 2
- 229930012538 Paclitaxel Natural products 0.000 claims description 2
- 229910001308 Zinc ferrite Inorganic materials 0.000 claims description 2
- 238000007664 blowing Methods 0.000 claims description 2
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 claims description 2
- 229960004316 cisplatin Drugs 0.000 claims description 2
- 229910052802 copper Inorganic materials 0.000 claims description 2
- 125000003916 ethylene diamine group Chemical group 0.000 claims description 2
- 238000001215 fluorescent labelling Methods 0.000 claims description 2
- 150000002500 ions Chemical class 0.000 claims description 2
- 229910052742 iron Inorganic materials 0.000 claims description 2
- 229910052748 manganese Inorganic materials 0.000 claims description 2
- 229910052759 nickel Inorganic materials 0.000 claims description 2
- 229960001592 paclitaxel Drugs 0.000 claims description 2
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 claims description 2
- 229910006297 γ-Fe2O3 Inorganic materials 0.000 claims description 2
- 238000003384 imaging method Methods 0.000 abstract description 4
- 108090000623 proteins and genes Proteins 0.000 abstract description 4
- 208000037260 Atherosclerotic Plaque Diseases 0.000 abstract description 2
- 239000002872 contrast media Substances 0.000 abstract description 2
- 230000015572 biosynthetic process Effects 0.000 abstract 2
- 206010028980 Neoplasm Diseases 0.000 abstract 1
- 229920001610 polycaprolactone Polymers 0.000 description 98
- 239000004632 polycaprolactone Substances 0.000 description 65
- LCGLNKUTAGEVQW-UHFFFAOYSA-N Dimethyl ether Chemical compound COC LCGLNKUTAGEVQW-UHFFFAOYSA-N 0.000 description 23
- 239000000463 material Substances 0.000 description 9
- 239000002105 nanoparticle Substances 0.000 description 8
- 229920000747 poly(lactic acid) Polymers 0.000 description 7
- CKLJMWTZIZZHCS-UHFFFAOYSA-N D-OH-Asp Natural products OC(=O)C(N)CC(O)=O CKLJMWTZIZZHCS-UHFFFAOYSA-N 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- CKLJMWTZIZZHCS-UWTATZPHSA-N L-Aspartic acid Natural products OC(=O)[C@H](N)CC(O)=O CKLJMWTZIZZHCS-UWTATZPHSA-N 0.000 description 6
- 229960005261 aspartic acid Drugs 0.000 description 6
- 238000010586 diagram Methods 0.000 description 6
- 239000004626 polylactic acid Substances 0.000 description 6
- 238000001228 spectrum Methods 0.000 description 6
- GMPKIPWJBDOURN-UHFFFAOYSA-N Methoxyamine Chemical group CON GMPKIPWJBDOURN-UHFFFAOYSA-N 0.000 description 5
- 238000000502 dialysis Methods 0.000 description 5
- 239000003999 initiator Substances 0.000 description 5
- 239000013067 intermediate product Substances 0.000 description 5
- 229920001427 mPEG Polymers 0.000 description 5
- 238000007151 ring opening polymerisation reaction Methods 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- 238000002296 dynamic light scattering Methods 0.000 description 4
- 238000012637 gene transfection Methods 0.000 description 3
- 239000010410 layer Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- UDOPJKHABYSVIX-UHFFFAOYSA-N 2-[4,7,10-tris(carboxymethyl)-6-[(4-isothiocyanatophenyl)methyl]-1,4,7,10-tetrazacyclododec-1-yl]acetic acid Chemical compound C1N(CC(O)=O)CCN(CC(=O)O)CCN(CC(O)=O)CCN(CC(O)=O)C1CC1=CC=C(N=C=S)C=C1 UDOPJKHABYSVIX-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 2
- 230000009920 chelation Effects 0.000 description 2
- 239000007795 chemical reaction product Substances 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 238000012377 drug delivery Methods 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 2
- 230000005311 nuclear magnetism Effects 0.000 description 2
- 229920000835 poly(gamma-benzyl-L-glutamate) polymer Polymers 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- IMTCOHJNTRYIGD-UHFFFAOYSA-N CNC1C=[C+]CC1 Chemical compound CNC1C=[C+]CC1 IMTCOHJNTRYIGD-UHFFFAOYSA-N 0.000 description 1
- 229910021592 Copper(II) chloride Inorganic materials 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 239000005977 Ethylene Substances 0.000 description 1
- 229910003317 GdCl3 Inorganic materials 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- FLSPFAWUTVGOAX-ZEQRLZLVSA-N [(2s)-6-amino-2-(phenylmethoxycarbonylamino)hexanoyl] (2s)-6-amino-2-(phenylmethoxycarbonylamino)hexanoate Chemical compound N([C@@H](CCCCN)C(=O)OC(=O)[C@H](CCCCN)NC(=O)OCC=1C=CC=CC=1)C(=O)OCC1=CC=CC=C1 FLSPFAWUTVGOAX-ZEQRLZLVSA-N 0.000 description 1
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- 125000003636 chemical group Chemical group 0.000 description 1
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- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
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Abstract
The invention discloses a kind of polyethylene glycol-amino acid-polyester three-block polymkeric substance and its production and use, be characterized in the carrier using the amphiphilic formation micella of polyethylene glycol-amino acid-polyester three-block polymkeric substance as gene, medicine and contrast medium, build multifunctional nano composite particles further.This triblock polymer Biocompatibility is good, and the nano-micelle middle layer of formation can be cross-linked, the nano-complex particle Stability Analysis of Structures of structure, energy functional modification further.The multifunctional nano composite particles built is used for atherosclerotic plaque development, tumor imaging or medicine controlled releasing.<!--1-->
Description
Technical Field
The invention relates to a Polyethylene glycol-Polyamino acid-Polyester (Polyethylene glycol-Polyamino acid-Polyester) triblock high-molecular polymer and a preparation method and application thereof, belonging to the fields of biological medicine materials, medicine delivery and molecular imaging.
Background
At present, multifunctional composite nanoparticles (such as photoresponse nanoparticles, magnetic response nanoparticles and electric response nanoparticles) are widely applied to the biological fields of molecular imaging, drug delivery, gene transfection, cell sorting and the like. There are many materials used for constructing multifunctional composite nanoparticles, such as dendrimers, inorganic nanoparticles, and amphiphilic block polymers. The amphiphilic block polymers are most widely used (Adv Drug Deliv Rev 1996;21(2):107-16), and most of the amphiphilic block polymers are diblock polymers, such as polyethylene glycol-polycaprolactone (J Control Release 2004;98(3):415-26), polyethylene glycol-polylactide (Langmuir 2003;19: 8428-. When the concentration of the amphiphilic block polymer is higher than the critical micelle concentration, nano-micelles are formed in aqueous solution through the hydrophilic and hydrophobic interaction, hydrophobic drugs and functional inorganic nano-particles are wrapped in hydrophobic cores of the micelles (Science 2002;298(5599): 1759-62), hydrophilic shells can complex genes (Adv Drug Deliv Rev 2002;54(2): 203-22), and functional small molecules (such as small molecule magnetic resonance contrast agents, fluorescent molecules, small molecules with radioactive labels or active polypeptides) are introduced through covalent bonds (Angew Chem Int EdEngl 2004;43(46): 6323-7). The stability of the nano micelle is enhanced, the critical micelle concentration of the amphiphilic block polymer is reduced, the drug delivery efficiency and the gene transfection efficiency of the nano composite particles can be improved, and the biomedical application of the material is facilitated. Therefore, the block copolymer is a hot spot for the research of amphiphilic block polymers.
Disclosure of Invention
The invention aims to provide a polyethylene glycol-polyamino acid-polyester triblock polymer and a preparation method and application thereof aiming at the defects of the prior art, and is characterized in that the amphiphilic polymer forms micelles to be used as carriers of genes, medicines and contrast agents, and multifunctional nano composite particles are further constructed. The multifunctional nano composite particle is applied to development of atherosclerotic plaques, tumor development and drug controlled release.
The aim of the invention is realized by the following technical measures, wherein the parts of the raw materials are mole parts except for special description.
The chemical structural formula of the polyethylene glycol-polyamino acid-polyester triblock polymer is as follows:
wherein,
R1 = -CH3、-CH2(CH2)0～5COOH、-CH2(CH2)0～5OH 、-CH2(CH2)0～5NH2、-CH2(CH2)0～5SH、-CH2(CH2)0～5N3、-CH2(CH2) 0～5C≡CH;
R2 = -CONHCH2CH2、-(CH2)1～5、-COCH2CH2CONHCH2CH2;
R3 = -CH2COOH、-CH2CH2COOH、-CH2CH2CH2CH2NH2;
R4 = -(CH2)1～5、-CH(CH3)
m, n and q are independently 10 to 500.
The preparation method of the polyethylene glycol-polyamino acid-polyester triblock polymer comprises the following steps:
(1) preparation of polyethylene glycol-polyamino acid two-block polymer
Under the protection of inert gas, weighing 0.1 molar part of dried polyethylene glycol with one end being amino, dissolving the polyethylene glycol in a reaction bottle by using 5-100 parts of anhydrous dichloromethane, weighing 0.5-10 molar parts of amino acid-N-cyclic carbonic anhydride, dissolving the amino acid-N-cyclic carbonic anhydride by using 0.5-10 parts of anhydrous N, N-dimethylformamide, then adding the mixture into the reaction bottle, reacting at the temperature of 20-50 ℃ for 12-72h, evaporating to remove the solvent, dissolving the product by using 5-30 parts of chloroform, precipitating in 50-300 parts of ethyl acetate, collecting the precipitate, dissolving in 5-30 parts of chloroform, precipitating in 50-300 parts of ethyl acetate, repeating for three times to obtain a polyethylene glycol-polyamino acid two-block polymer;
wherein, the inert gas is any one of argon, nitrogen or helium; the amino acid is any one of L-glutamic acid-gamma-benzyl ester, L-aspartic acid-beta-benzyl ester, O-benzyl-L-tyrosine or N-benzyloxycarbonyl-L-lysine;
(2) preparation of polyethylene glycol-polyamino acid-polyester triblock polymer
Weighing 0.1 molar part of polyethylene glycol-polyamino acid diblock polymer and 0.5-10 molar parts of polyester monomer in a reaction bottle, adding 5-50 parts of anhydrous dioxane, toluene or benzyl ether for dissolving under the protection of inert gas, adding 0.0001-0.1 molar part of stannous octoate, placing the reaction bottle at the temperature of 105 ℃ and 160 ℃ for reacting for 12-72h, evaporating to remove the solvent, dissolving the product with 5-30 parts of chloroform, precipitating in 50-300 parts of glacial ethyl ether, collecting the precipitate, dissolving in 5-30 parts of chloroform, precipitating in 50-300 parts of glacial ethyl ether, repeating for three times to obtain the polyethylene glycol-polyamino acid-polyester triblock polymer;
wherein, the inert gas is any one of argon, nitrogen or helium; the polyester monomer is caprolactone or lactide;
(3) polyethylene glycol-polyamino acid-polyester triblock polymer de-side group protection
Adding 0.1 molar part of polyethylene glycol-polyamino acid-polyester triblock polymer into a high-pressure reaction kettle, dissolving 100 parts of tetrahydrofuran, adding 0.8% palladium carbon, introducing hydrogen with 2-10 atmospheric pressures, reacting at the temperature of 30-50 ℃ for 24-72h, filtering to remove the palladium carbon, and performing rotary evaporation to remove the tetrahydrofuran to obtain the side group-removed polyethylene glycol-polyamino acid-polyester triblock polymer.
One of the uses of the polyethylene glycol-polyamino acid-polyester triblock polymer
(1) Preparation of polyethylene glycol-polyamino acid-polyester triblock polymer nano micelle
Dissolving a polyethylene glycol-polyamino acid-polyester triblock polymer in an organic solvent, preparing a solution with the concentration of 5-20mg/ml, taking 0.5-10 parts of the solution, dripping the solution into 5-100 parts of an aqueous phase solvent at the temperature of 25-30 ℃ and the ultrasonic power of 130W and the amplitude of 65 percent, wherein the aqueous phase solvent is pure water or a phosphoric acid buffer solution with the pH value of 4-12, and shaking up; removing residual organic solvent in the aqueous phase solution by a reduced pressure rotary evaporation method to prepare the polyethylene glycol-polyamino acid-polyester triblock polymer nano micelle;
wherein the organic solvent is any one of chloroform, tetrahydrofuran or ethanol;
(2) polyethylene glycol-polyamino acid-polyester triblock polymer micelle loaded nanocrystal
a. Adding 2-10mg/ml single nano crystal dispersed in organic phase into a reagent bottle, removing organic solvent by vacuum drying or blowing dry by inert gas, and then weighing 0.001-0.5 part of nano crystal;
b. dissolving 0.005-1 part of triblock polymer in 0.5-10 parts of organic phase solvent, adding the solution into the nanocrystal, and uniformly mixing by shaking; adding the organic solution into 5-100 parts of water phase solvent at 25-30 ℃ and ultrasonic power of 130W and amplitude of 65%, wherein the water phase solvent is pure water or phosphoric acid buffer solution with pH value of 4-12, and shaking up; removing residual organic solvent in the aqueous phase solution by a reduced pressure rotary evaporation method to prepare nano micelle of the triblock polymer coated nanocrystal;
wherein, the organic phase solvent is any one of dichloromethane, trichloromethane, tetrahydrofuran or ethanol; the nanocrystal is Fe3O4Nanocrystalline, gamma-Fe2O3Nanocrystals, ZnFe2O4Nanocrystals, MnFe2O4Nanocrystals, CoFe2O4Any one of nanocrystals, FePt nanocrystals, Au nanocrystals, Ag nanocrystals, CdSe nanocrystals, CdZnS nanocrystals, CdSe/ZnS nanocrystals, or CdSe/CdS nanocrystals;
(3) polyethylene glycol-polyamino acid-polyester triblock polymer micelle loaded with hydrophobic drug
Weighing 0.001-0.5 part of hydrophobic drug and 0.005-1 part of triblock polymer, mixing and dissolving into 0.5-10 parts of organic phase solvent, uniformly mixing, adding into 5-100 parts of aqueous phase solvent at 25-30 ℃, ultrasonic power of 130W and amplitude of 65%, wherein the aqueous phase solvent is pure water or phosphoric acid buffer solution with pH value of 4-12, and shaking uniformly; removing residual organic solvent in the aqueous phase solution by a reduced pressure rotary evaporation method to prepare the nano micelle of the triblock polymer coated drug;
wherein, the organic phase solvent is any one of trichloromethane, tetrahydrofuran or ethanol; the hydrophobic drug is any one of adriamycin, paclitaxel or cisplatin;
use two of said polyethylene glycol-polyamino acid-polyester triblock polymer
1) Micellar internal crosslinking of polyethylene glycol-polyamino acid-polyester triblock polymer micelles
Adding 0.005-0.05 part of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride into 5 parts of polyethylene glycol-polyamino acid-polyester triblock polymer micelle solution with the concentration of 1-10mg/ml, reacting at room temperature for 10-30min, adding 0.001-0.01 part of cross-linking agent, stirring overnight, and dialyzing for 12-24h to obtain the polyethylene glycol-polyamino acid-polyester triblock polymer micelle with the cross-linked amino acid layer;
2) multifunctional nano composite particle is constructed by taking polyethylene glycol-polyamino acid-polyester triblock polymer micelle as platform
(1) The surface of micelle particles prepared by triblock polymer is modified by chemical covalent bonds to introduce micromolecular ligands, and the micelle particles are further chelated with metal ions for magnetic resonance or nuclide development;
wherein, the chemical covalent bond modification is performed by condensation reaction of carboxyl and amino, esterification reaction of carboxyl and hydroxyl, click reaction of alkynyl and azide, reaction of halide and amino, and reaction of amino and aldehyde group;
(2) introducing fluorescent molecules to the surfaces of micelle particles prepared by triblock polymers through chemical modification for fluorescent labeling;
wherein, the chemical modification is carried out by condensation reaction of carboxyl and amino, esterification reaction of carboxyl and hydroxyl, click reaction of alkynyl and azide, reaction of halide and amino, and reaction of amino and aldehyde group;
(3) the surface of micelle particles prepared by triblock polymer is introduced with functional short peptide for cell or tissue targeting through chemical modification;
wherein, the chemical modification is carried out by condensation reaction of carboxyl and amino, esterification reaction of carboxyl and hydroxyl, click reaction of alkynyl and azide, reaction of halide and amino, and reaction of amino and aldehyde group; the accessed functional short peptide is: arginine-glycine-aspartic acid or a cell-penetrating peptide.
The cross-linking agent is ethylenediamine or 1, 4-butanediamine.
The small molecular ligand is any one of 1,4,7, 10-tetraazacyclododecane 1,4,7, 10-tetraacetic acid, 1,4,7, 10-tetraazacyclododecane N, N' -triacetic acid or diethyltriaminepentaacetic acid.
The metal ions are: ions with +2 or +3 valence in Mn, Fe, Co, Ni, Cu, Ga, Tc and Gd.
Performance testing
1. Characterizing intermediate products and final products in the process of preparing the triblock polymer by adopting a hydrogen nuclear magnetic spectrum; as shown in detail in FIGS. 1-3 and 6-8, the results show that the obtained product is consistent with the target product according to the one-to-one correspondence of different hydrogen atoms in the product molecule and the peak area and the displacement in the spectrogram.
2. The internal crosslinking of the formed triblock polymer nano-micelle is characterized by adopting a scanning electron microscope; as shown in fig. 4A and fig. 4B, after freeze-drying the micelle before and after crosslinking, respectively, the micelle is dissolved in an organic solvent, and the micelle before crosslinking is dispersed by observation of a scanning electron microscope, while the micelle after crosslinking still maintains a spherical state.
3. Adopting a potential particle size tester to test the particle size of the tri-block polymer nano micelle loaded with ferroferric oxide; as shown in detail in fig. 5.
4. Adopting a potential particle size tester to test the particle size and potential of the formed triblock polymer nano micelle; this is shown in detail in figure 9.
The invention has the following advantages:
1 the triblock polymer consists of polyamino acid, polyester and polyethylene glycol, and has good biocompatibility.
2 preparing the polyethylene glycol-polyamino acid-polyester triblock amphiphilic polymer through two-step polymerization reaction.
3 the triblock amphiphilic polymer can wrap the nano-crystal and the hydrophobic drug, so that the nano-crystal and the hydrophobic drug are transferred from an organic phase to a water phase, the original physical properties of the nano-crystal are not changed after the nano-crystal and the hydrophobic drug are transferred to the water phase, and the particle size of the prepared nano-composite particle is adjustable.
4 the nano micelle interlayer formed by the triblock polymer can be crosslinked, and the constructed nano composite particle has a stable structure and can be further functionally modified.
5 the nano composite particle constructed by the triblock amphiphilic polymer can be used for compounding genes and used as a gene transfection vector.
6 the surface of the nano composite particle constructed by the triblock amphiphilic polymer can be modified by various chemical groups, and image molecules such as optical, magnetic resonance or nuclide and the like are introduced, so that the nano composite particle can be used for marking cells and can be used for in-vivo multifunctional development imaging.
Drawings
FIG. 1 is a schematic diagram of the preparation of methyl-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone (CH)3PEG-PASP-PCL), the intermediate product of the hydrogen nuclear magnetic spectrum of the end methyl ether polyethylene glycol-poly-L-aspartic acid-beta-benzyl ester.
FIG. 2 is a schematic diagram of the preparation of methyl-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone (CH)3PEG-PASp-PCL), intermediate product terminal methyl ether polyHydrogen nuclear magnetic spectrum of ethylene glycol-poly-L-aspartic acid-beta-benzyl-polycaprolactone.
FIG. 3 is a schematic diagram of the preparation of methyl-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone (CH)3PEG-PASp-PCL), the final product, end methyl ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone.
FIG. 4A terminal methyl ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone (CH)3-PEG-PASp-PCL) micelles in chloroform.
FIG. 4B terminal methyl ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone (CH)3-PEG-PASp-PCL) micelle is dissolved in chloroform after being crosslinked for scanning electron microscope picture.
FIG. 5 shows the particle size distribution of the nanocomposite particles constructed by wrapping Fe3O4 nanocrystals with PEG-poly-L-aspartic acid-polycaprolactone (dynamic light scattering method).
FIG. 6 is a schematic diagram of the preparation of amino terminated poly (ethylene glycol) -poly (L-aspartic acid) -polycaprolactone (NH)2PEG-PASp-PCL), the intermediate product has a hydrogen nuclear magnetic spectrum of nitrogen-terminated and nitrogen-dibenzyl polyethylene glycol-poly-L-aspartic acid-beta-benzyl ester.
FIG. 7 is a schematic diagram of the preparation of amino terminated poly (ethylene glycol) -poly (L-aspartic acid) -polycaprolactone (NH)2-PEG-PASp-PCL), the intermediate product has a hydrogen nuclear magnetic spectrum of N, N-dibenzyl polyethylene glycol-poly-L-aspartic acid-beta-benzyl ester-polycaprolactone.
FIG. 8 is a schematic diagram of the preparation of amino terminated poly (ethylene glycol) -poly (L-aspartic acid) -polycaprolactone (NH)2PEG-PASp-PCL), the final product amino polyethylene glycol-poly-L-aspartic acid-polycaprolactone hydrogen nuclear magnetic spectrum.
Fig. 9 shows the particle size distribution of the peg-poly-L-aspartic acid-polycaprolactone nanomicelle (dynamic light scattering).
Detailed Description
The present invention is described in detail below by way of examples, it should be noted that the examples are only for the purpose of further illustration, and are not to be construed as limiting the scope of the present invention, and that those skilled in the art can make insubstantial modifications and adaptations of the present invention based on the teachings of the present invention described above.
Example 1: methyl-terminated ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone (CH)3-PEG-PASp-PCL) preparation steps and technological parameters
The chemical reaction formula is as follows:
the preparation steps and the technological parameters are as follows:
1) terminal methyl ether polyethylene glycol-poly-L-aspartic acid-beta-benzyl ester (CH)3PEG-PBLA) preparation:
polyethylene glycol-poly beta-benzyl-L-aspartic acid adopts amino-terminated monomethyl ether polyethylene glycol (CH)3-PEG-NH2) The initiator is N-carboxylic anhydride (OBzl-Asp-NCA) ring-opening polymerization of L-aspartic acid-beta-benzyl ester. First, 0.1mmol of dried terminal amino methyl ether polyethylene glycol (CH) was precisely weighed3-PEG-NH2Molecular weight 5000 g/mol) in a reaction bottle, and adding 5ml of anhydrous dichloromethane under the protection of argon to dissolve. Then 0.5 mmol of N-carboxyanhydride of L-aspartic acid-beta-benzyl ester (OBzl-Asp-NCA) is accurately weighed, dissolved by 0.5 ml of anhydrous N, N-Dimethylformamide (DMF) under the protection of argon and added into a reaction bottle. The reaction flask was placed in a 20 ℃ oil bath for 12 hours. After the reaction was completed, the solvent was evaporated, dissolved in 5ml of chloroform and precipitated with 50ml of glacial ethyl ether. The precipitate was collected, redissolved in 5ml chloroform, precipitated with 50ml of ethyl acetate and repeated three times. The precipitate was dried in vacuo to give the product as a white solid.
2) Methyl-terminated ether polyethylene glycol-poly (L-aspartic acid) -beta-benzyl-polycaprolactone (CH)3Preparation of PEG-PBLA-PCL):
0.1mmol of polyethylene glycol-poly-L-aspartic acid-beta-benzyl ester (CH) is weighed in a reaction bottle3PEG-PBLA) and 0.5 mmol caprolactone monomer, 5ml of anhydrous toluene were added under argon protection. After all the materials are dissolved, adding 0.0001 mmol of stannous octoate under the protection of argon, and placing the reaction bottle in an oil bath at 105 ℃ for reaction for 12 hours. After the reaction was completed, the solvent was evaporated, dissolved in 5ml of chloroform and precipitated with 50ml of glacial ethyl ether. The precipitate was collected, dissolved in 5ml of chloroform and precipitated with 50ml of ethyl acetate and repeated three times. The precipitate was dried in vacuo to give the product as a white solid.
3) Methyl-terminated ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone (CH)3Preparation of PEG-PASp-PCL):
0.1mmol of polyethylene glycol-poly beta-benzyl-L-aspartic acid-polycaprolactone is dissolved in 100ml of tetrahydrofuran, 8% of palladium carbon is added, and the mixture is added into a high-pressure reaction kettle. Introducing 2 atmospheres of hydrogen, and reacting at 30 ℃ for 24 hours. After the reaction is finished, filtering to remove palladium carbon, and performing rotary evaporation to remove tetrahydrofuran.
As shown in FIGS. 1-3, the reaction product of each step was characterized by hydrogen nuclear magnetism.
Example 2: methyl-terminated ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone (CH)3-preparation of PEG-PAsp-PCL) micelles and amino acid layer cross-linking:
dissolving 3 mg of methyl ether terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone in 0.5 ml of tetrahydrofuran, adding the solution into 5 g of pure water at the temperature of 25 ℃, with the ultrasonic power of 130W and the amplitude of 65%, and shaking up for 24 hours; removing residual tetrahydrofuran solvent in the aqueous phase solution by reduced pressure rotary evaporation method, and concentrating to 3 ml; the hollow micelle of the end methyl ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone is prepared. The particle size distribution measured by dynamic light scattering is shown in FIG. 9.
3 mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride was weighed, added to the above 3ml of the micelle solution, and reacted at room temperature for 10 minutes. Adding 0.6 mg of 1, 4-butanediamine and stirring overnight; to obtain the amino acid layer cross-linked terminal methyl ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone (CH)3-PEG-PAsp-PCL) micelles. As shown in fig. 4B, the crosslinked micelle is dissolved in an organic solvent after being lyophilized, and still maintains a spherical state.
Example 3: methyl-terminated ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone (CH)3PEG-PASp-PCL) for coating ferroferric oxide nano-particles
Adding the Fe3O4 nanocrystals with the concentration of 10mg/ml and single dispersion in n-hexane into a reagent bottle, removing the organic solvent in a vacuum drying mode, and then weighing 1mg of the Fe3O4 nanocrystals; dissolving 3 mg of methyl ether terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone in 0.5 ml of tetrahydrofuran, and uniformly mixing the solution with the dried nanocrystal at 25 ℃ by shaking; adding the organic solution into 5 g of pure water at the temperature of 25 ℃, the ultrasonic power of 130W and the amplitude of 65%, and shaking up for 24 hours; removing residual tetrahydrofuran solvent in the aqueous phase solution by a reduced pressure rotary evaporation method; preparing the nano composite particles constructed by the Fe3O4 nano crystal wrapped by the methyl-terminated ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone. The particle size distribution of the particles obtained by dynamic light scattering is shown in FIG. 5.
Example 4: methyl-terminated ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone (CH)3PEG-PASp-PCL) for encapsulating hydrophobic doxorubicin
Dissolving 3 mg of methyl ether terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone in 0.5 ml of tetrahydrofuran, adding the solution into a reagent bottle filled with 1mg of hydrophobic adriamycin, and shaking and mixing the solution uniformly at the temperature of 25 ℃; adding the organic solution into 5 g of pure water at the temperature of 25 ℃, the ultrasonic power of 130W and the amplitude of 65%, and shaking up for 24 hours; removing residual tetrahydrofuran solvent in the aqueous phase solution by a reduced pressure rotary evaporation method; preparing the nano composite particles constructed by the Fe3O4 nano crystal wrapped by the methyl-terminated ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone.
Example 5: methyl-terminated ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone (CH)3PEG-PASp-PCL) for coating CdSe/ZnS nanocrystals
Adding the CdSe/ZnS nanocrystals with the concentration of 4 mg/ml and mono-dispersed in chloroform into a reagent bottle, removing an organic solvent in a vacuum drying mode, and then weighing 2 mg of the CdSe/ZnS nanocrystals; uniformly mixing 1 ml of tetrahydrofuran solution of terminal methyl ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone with the concentration of 6 mg/ml with the dried nanocrystal at the temperature of 25 ℃ by shaking; adding the organic solution into 10 g of pure water at the temperature of 25 ℃, the ultrasonic power of 130W and the amplitude of 65%, and shaking up for 24 hours; removing residual tetrahydrofuran solvent in the aqueous phase solution by a reduced pressure rotary evaporation method; the nano composite particle constructed by CdSe/ZnS nano crystal wrapped by methyl-terminated ether polyethylene glycol-poly-L-aspartic acid-polycaprolactone is prepared.
Example 6: amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone (NH)2-PEG-PASp-PCL) preparation steps and technological parameters
The chemical reaction formula is as follows:
the preparation steps and the technological parameters are as follows:
1) n-terminated, N-dibenzyl polyethylene glycol-poly-L-aspartic acid-beta-benzyl ester (Bn)2-preparation of N-PEG-PBLA):
firstly, accurately weighing 0.1mmol of dry terminal nitrogen and nitrogen dibenzyl polyethylene glycol (Bn)2-N-PEG-NH24600 g/mol) was placed in a reaction flask and dissolved by adding 100ml of anhydrous dichloromethane under the protection of argon. Then 10 mmol of N-carboxyanhydride (OBzl-Asp-NCA) of L-aspartic acid-beta-benzyl ester is accurately weighed, dissolved by 10 ml of anhydrous N, N-Dimethylformamide (DMF) under the protection of argon and added into a reaction bottle, the reaction bottle is placed in an oil bath at 50 ℃ for reaction for 72 hours, after the reaction is finished, the solvent is evaporated and removed, and then dissolved in 30ml of chloroform and precipitated by 300ml of glacial ethyl ether. The precipitate was collected, dissolved in 30ml of chloroform and precipitated with 300ml of glacial ethyl ether, which was repeated three times. The precipitate was dried in vacuo to give a white product.
2) N-terminated, N-dibenzylpolyethylene glycol-poly (L-aspartic acid) -beta-benzyl-polycaprolactone (Bn)2-preparation of N-PEG-PBLA-PCL):
0.1mmol of end nitrogen and nitrogen dibenzyl polyethylene glycol-poly-L-aspartic acid-beta-benzyl ester (Bn) is weighed in a reaction bottle2-N-PEG-PBLA) and 10 mmol caprolactone monomer, 50ml anhydrous benzyl ether was added under nitrogen. After all the materials are dissolved, adding 0.1mmol of stannous octoate under the protection of argon, and placing the reaction bottle in an oil bath at 160 ℃ for reaction for 72 hours. After the reaction was completed, the solvent was evaporated, dissolved in 30ml of chloroform, and precipitated with 300ml of glacial ethyl ether. The precipitate was collected, dissolved in 30ml of chloroform and precipitated with 300ml of glacial ethyl ether, which was repeated three times. The precipitate was dried in vacuo to give a white product.
3) Preparation of amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone (NH 2-PEG-PASp-PCL):
0.1mmol of terminal nitrogen and nitrogen dibenzyl polyethylene glycol-poly beta-benzyl-L-aspartic acid-polycaprolactone is dissolved in 400ml of tetrahydrofuran, 8% of palladium carbon is added, and the mixture is added into a high-pressure reaction kettle. Hydrogen gas of 10 atm was introduced, and the reaction was carried out at 50 ℃ for 72 hours. After the reaction is finished, filtering to remove palladium carbon, and performing rotary evaporation to remove tetrahydrofuran.
As shown in FIGS. 6-8, the reaction product of each step was characterized by hydrogen nuclear magnetism.
Example 7: 1,4,7, 10-tetraazacyclododecane 1,4,7, 10-tetraacetic acid is introduced to the surface of amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone (NH2-PEG-PASp-PCL) nano micelle particles and is further mixed with Gd3+Chelation for magnetic resonance imaging
Dissolving 3 mg of amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone in 0.5 ml of tetrahydrofuran, adding the mixture into 5 g of pure water at the temperature of 25 ℃, with the ultrasonic power of 130W and the amplitude of 65%, and shaking up for 24 hours; removing residual tetrahydrofuran solvent in the aqueous phase solution by a reduced pressure rotary evaporation method; 3ml of amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone micelle is prepared.
1mg of 2- (4-isothiocyanatobenzyl) -1,4,7, 10-tetraazacyclododecane 1,4,7, 10-tetraacetic acid was dissolved in 0.5 ml of dimethyl sulfoxide, added dropwise to the amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone micelle solution, and stirred at room temperature overnight. Dialyzing the solution with 1 ten thousand molecular weight dialysis bag for 2 days, adding 1mg GdCl3The solution was stirred overnight to adjust the pH to 6.5. Dialyzing in dialysis bag with molecular weight of 1 ten thousand for 2 days to obtain magnetic resonance T1Developing the effect of the nanomicelle.
Example 8: introducing 1,4,7, 10-tetraazacyclododecane 1,4,7, 10-tetraacetic acid on the surface of amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone (NH2-PEG-PASp-PCL) nano micelle particle, and further reacting with the amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone (NH2-PEG-PASp-PCL) nano micelle particle64Cu chelation for nuclide development
Dissolving 3 mg of amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone in 0.5 ml of tetrahydrofuran, adding the mixture into 5 g of pure water at the temperature of 25 ℃, with the ultrasonic power of 130W and the amplitude of 65%, and shaking up for 24 hours; removing residual tetrahydrofuran solvent in the aqueous phase solution by a reduced pressure rotary evaporation method; 3ml of amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone micelle is prepared.
1mg of 2- (4-isothiocyanatobenzyl) -1,4,7, 10-tetraazacyclododecane 1,4,7, 10-tetraacetic acid was dissolved in 0.5 ml of dimethyl sulfoxide, added dropwise to the amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone micelle solution, and stirred at room temperature overnight. The solution was dialyzed for 2 days using a dialysis bag of 1 ten thousand molecular weight. 50 ul of the solution was added64CuCl210 mM hydrochloric acid solution (2.35 mCi), the pH of the solution being adjusted to 8. Incubating at 40 ℃ for 20 minutes to obtain64Cu-labeled nanomicelles.
Example 9: introducing a fluorescent label on the surface of the amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone (NH2-PEG-PASp-PCL) nano micelle particle
Dissolving 3 mg of amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone in 0.5 ml of tetrahydrofuran, adding the mixture into 5 g of pure water at the temperature of 25 ℃, with the ultrasonic power of 130W and the amplitude of 65%, and shaking up for 24 hours; removing residual tetrahydrofuran solvent in the aqueous phase solution by a reduced pressure rotary evaporation method; 3ml of amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone micelle is prepared.
Dissolving 1mg fluorescein isothiocyanate in 0.5 ml dimethyl sulfoxide, dripping into amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone micelle solution, and stirring at room temperature overnight. Dialyzing the solution for 2 days by using a dialysis bag with the molecular weight of 1 ten thousand to obtain the nano micelle marked by the fluorescein.
Example 10: targeting short peptide is introduced to the surface of amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone (NH2-PEG-PASp-PCL) nano micelle particle
Dissolving 3 mg of amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone in 0.5 ml of tetrahydrofuran, adding the mixture into 5 g of pure water at the temperature of 25 ℃, with the ultrasonic power of 130W and the amplitude of 65%, and shaking up for 24 hours; removing residual tetrahydrofuran solvent in the aqueous phase solution by a reduced pressure rotary evaporation method; 3ml of amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone micelle is prepared.
1mg of carboxyl-activated arginine-glycine-aspartic acid is dissolved in 0.5 ml of N, N-dimethylformamide, added dropwise into the amino-terminated polyethylene glycol-poly-L-aspartic acid-polycaprolactone micelle solution, and stirred at room temperature overnight. Dialyzing the solution by a dialysis bag with the molecular weight of 1 ten thousand for 2 days to obtain the nano micelle with the surface introduced with the arginine-glycine-aspartic acid.
Example 11: preparation steps and process parameters of methyl-terminated ether polyethylene glycol-poly-L-glutamic acid-polycaprolactone (CH3-PEG-PLGA-PCL)
The chemical reaction formula is as follows:
the preparation steps and the technological parameters are as follows:
1) terminal methyl ether polyethylene glycol-poly-L-glutamic acid-gamma-benzyl ester (CH)3-preparation of PEG-PBLG):
polyethylene glycol-poly beta-benzyl-L-aspartic acid adopts amino-terminated monomethyl ether polyethylene glycol (CH)3-PEG-NH2) The initiator is N-carboxylic anhydride ring-opening polymerization of L-glutamic acid-gamma-benzyl ester. First, 0.1mmol of dried terminal amino methyl ether polyethylene glycol (CH) was precisely weighed3-PEG-NH2Molecular weight 5000 g/mol) in a reaction bottle, and adding 20ml of anhydrous dichloromethane to dissolve under the protection of argon. Then 3 mmol of N-carboxylic anhydride of L-glutamic acid-gamma-benzyl ester is accurately weighed, dissolved by 2ml of anhydrous N, N-Dimethylformamide (DMF) under the protection of argon and added into a reaction bottle. The reaction flask was placed in a 35 ℃ oil bath for 72 hours. After the reaction was completed, the solvent was evaporated, dissolved in 30ml of chloroform, and precipitated with 300ml of glacial ethyl ether. The precipitate was collected, dissolved in 30ml of chloroform,precipitation with 300ml of glacial ethyl ether was repeated three times. The precipitate was dried in vacuo to give a white product.
2) Methyl-terminated ether polyethylene glycol-poly-L-glutamic acid-gamma-benzyl ester-polycaprolactone (CH)3Preparation of PEG-PBLG-PCL):
0.1mmol of polyethylene glycol-poly-L-glutamic acid-gamma-benzyl ester (CH) is weighed in a reaction bottle3PEG-PBLG) and 4 mmol caprolactone monomer, 20ml of anhydrous toluene was added under argon protection. After all the materials are dissolved, adding 0.02mmol of stannous octoate under the protection of argon, and placing the reaction bottle in an oil bath at 110 ℃ for reaction for 48 hours. After the reaction was completed, the solvent was evaporated, dissolved in 30ml of chloroform, and precipitated with 300ml of glacial ethyl ether. The precipitate was collected, dissolved in 30ml of chloroform and precipitated with 300ml of glacial ethyl ether, which was repeated three times. The precipitate was dried in vacuo to give a white product.
3) Methyl-terminated ether polyethylene glycol-poly-L-glutamic acid-polycaprolactone (CH)3-preparation of PEG-PLGA-PCL):
0.1mmol of polyethylene glycol-poly-L-glutamic acid-gamma-benzyl ester-polycaprolactone is dissolved in 350ml of tetrahydrofuran, 8% of palladium carbon is added, and the mixture is added into a high-pressure reaction kettle. Hydrogen gas of 4 atm was introduced, and the reaction was carried out at 40 ℃ for 72 hours. After the reaction is finished, filtering to remove palladium carbon, and performing rotary evaporation to remove tetrahydrofuran.
Example 12: methyl-terminated ether polyethylene glycol-poly-L-lysine-polycaprolactone (CH)3-PEG-PLys-PCL) preparation steps and technological parameters
The chemical reaction formula is as follows:
the preparation steps and the technological parameters are as follows:
1) methyl-terminated ether polyethylene glycol-poly (N-benzyloxycarbonyl-L-lysine) (CH)3-PEG-PLys(Z)）The preparation of (1):
polyethylene glycol-poly beta-benzyl-L-aspartic acid adopts amino-terminated monomethyl ether polyethylene glycol (CH)3-PEG-NH2) The initiator is N-carboxylic anhydride ring-opening polymerization of L-glutamic acid-gamma-benzyl ester. First, 0.1mmol of dried terminal amino methyl ether polyethylene glycol (CH) was precisely weighed3-PEG-NH2Molecular weight 5000 g/mol) in a reaction bottle, and adding 20ml of anhydrous dichloromethane to dissolve under the protection of argon. Then 3 mmol of N-benzyloxycarbonyl lysine anhydride (Lys (z) -NCA) was precisely weighed, dissolved in 2ml of anhydrous N, N-Dimethylformamide (DMF) under an argon atmosphere, and then added to the reaction flask. The reaction flask was placed in a 35 ℃ oil bath for 72 hours. After the reaction was completed, the solvent was evaporated, dissolved in 30ml of chloroform, and precipitated with 300ml of glacial ethyl ether. The precipitate was collected, dissolved in 30ml chloroform, and precipitated with 300ml of glacial ethyl ether, repeated three times, and repeated three times.
2) Methyl-terminated ether polyethylene glycol-poly (N-benzyloxycarbonyl-L-lysine) -polycaprolactone (CH)3Preparation of PEG-PLys (Z) -PCL):
0.1mmol of polyethylene glycol-poly (N-benzyloxycarbonyl-L-lysine) (CH) is weighed in a reaction bottle3PEG-PLys (Z)) and 2mmol caprolactone monomer, 20ml anhydrous dioxane was added under argon. After all the materials are dissolved, adding 0.001mol of stannous octoate under the protection of argon, and placing the reaction bottle in an oil bath at 110 ℃ for reaction for 48 hours. After the reaction was completed, the solvent was evaporated, dissolved in 30ml of chloroform, and precipitated with 300ml of glacial ethyl ether. The precipitate was collected, dissolved in 30ml chloroform, and precipitated with 300ml of glacial ethyl ether, repeated three times, and repeated three times. The precipitate was dried in vacuo to give a white product.
3) Methyl-terminated ether polyethylene glycol-poly-L-lysine-polycaprolactone (CH)3-preparation of PEG-PLys-PCL):
0.1mmol of polyethylene glycol-poly (N-benzyloxycarbonyl-L-lysine) -polycaprolactone was dissolved in 400ml of tetrahydrofuran, and 8% palladium on carbon was added thereto together in a high-pressure reaction vessel. Hydrogen gas of 4 atm was introduced, and the reaction was carried out at 40 ℃ for 72 hours. After the reaction is finished, filtering to remove palladium carbon, and performing rotary evaporation to remove tetrahydrofuran.
Example 13: preparation steps and process parameters of terminal methyl ether polyethylene glycol-poly-L-aspartic acid-polylactate (CH3-PEG-PASp-PLA)
The chemical reaction formula is as follows:
the preparation steps and the technological parameters are as follows:
1) terminal methyl ether polyethylene glycol-poly-L-aspartic acid-beta-benzyl ester (CH)3PEG-PBLA) preparation:
polyethylene glycol-poly beta-benzyl-L-aspartic acid adopts amino-terminated monomethyl ether polyethylene glycol (CH)3-PEG-NH2) The initiator is N-carboxylic anhydride (OBzl-Asp-NCA) ring-opening polymerization of L-aspartic acid-beta-benzyl ester. First, 0.1mmol of dried terminal amino methyl ether polyethylene glycol (CH) was precisely weighed3-PEG-NH2Molecular weight 5000 g/mol) in a reaction bottle, and adding 20ml of anhydrous dichloromethane to dissolve under the protection of argon. Then 3 mmol of N-carboxyanhydride of L-aspartic acid-beta-benzyl ester (OBzl-Asp-NCA) is accurately weighed, dissolved by 2ml of anhydrous N, N-Dimethylformamide (DMF) under the protection of argon and added into a reaction bottle. The reaction flask was placed in a 35 ℃ oil bath for 72 hours. After the reaction was completed, the solvent was evaporated, dissolved in 30ml of chloroform, and precipitated with 300ml of glacial ethyl ether. The precipitate was collected, dissolved in 30ml of chloroform and precipitated with 300ml of glacial ethyl ether, which was repeated three times. The precipitate was dried in vacuo to give a white product.
2) Methyl-terminated ether polyethylene glycol-poly (L-aspartic acid) -beta-benzyl-polylactic acid (CH)3-PEG-PBLA-PLA) preparation:
0.1mmol of polyethylene glycol-poly L-aspartic acid-beta-benzyl ester (beta-benzyl ester) is weighed in a reaction bottleCH3PEG-PBLA) and 2mmol of lactide monomer, 20ml of anhydrous dioxane was added under argon protection. After all the materials are dissolved, adding 0.01 stannous octoate under the protection of argon, and placing the reaction bottle in an oil bath at 110 ℃ for reaction for 48 hours. After the reaction was completed, the solvent was evaporated, dissolved in 30ml of chloroform, and precipitated with 300ml of glacial ethyl ether. The precipitate was collected, dissolved in 30ml of chloroform and precipitated with 300ml of glacial ethyl ether, which was repeated three times. The precipitate was dried in vacuo to give a white product.
3) Methyl-terminated ether polyethylene glycol-poly-L-aspartic acid-polylactic acid (CH)3-preparation of PEG-PAsp-PLA):
0.1mmol of polyethylene glycol-poly beta-benzyl-L-aspartic acid-polylactic acid is dissolved in 400ml of tetrahydrofuran, 8 percent of palladium carbon is added, and the mixture is added into a high-pressure reaction kettle. Hydrogen gas of 4 atm was introduced, and the reaction was carried out at 40 ℃ for 72 hours. After the reaction is finished, filtering to remove palladium carbon, and performing rotary evaporation to remove tetrahydrofuran.
Example 14: methyl-terminated ether polyethylene glycol-poly-L-tyrosine-polycaprolactone (CH)3-PEG-PTyr-PCL) preparation steps and technological parameters
The chemical reaction formula is as follows:
the preparation steps and the technological parameters are as follows:
1) terminal methyl ether polyethylene glycol-poly-O-benzyl-L-tyrosine (CH)3-PEG-PBTyr) preparation:
polyethylene glycol-poly O-benzyl-L-tyrosine adopts amino-terminated monomethyl ether polyethylene glycol (CH)3-PEG-NH2) The initiator is N-carboxylic anhydride (OBzl-Tyr-NCA) of O-benzyl-L-tyrosine prepared by a ring-opening polymerization method. First, 0.1mmol of dried terminal amino methyl ether polyethylene glycol (CH) was precisely weighed3-PEG-NH2Molecular weight 5000 g/mol) in a reaction bottle, and adding 20ml of anhydrous dichloromethane to dissolve under the protection of argon. 3 mmol of OBzl-Tyr-NCA is accurately weighed, dissolved by 2ml of anhydrous N, N-Dimethylformamide (DMF) under the protection of argon and added into a reaction bottle. The reaction flask was placed in a 35 ℃ oil bath for 72 hours. After the reaction was completed, the solvent was evaporated, dissolved in 30ml of chloroform, and precipitated with 300ml of glacial ethyl ether. The precipitate was collected, dissolved in 30ml of chloroform and precipitated with 300ml of glacial ethyl ether, which was repeated three times. The precipitate was dried in vacuo to give a white product.
2) Methyl-terminated ether polyethylene glycol-poly-O-benzyl-L-tyrosine-polycaprolactone (CH)3Preparation of PEG-PBTyr-PCL):
0.1mmol of terminal methyl ether polyethylene glycol-poly O-benzyl-L-tyrosine (CH) is weighed in a reaction bottle3PEG-PBTyr) and 2.5mmol caprolactone monomer, under the protection of argon, were added to 20ml of anhydrous toluene. After all the materials are dissolved, adding a catalytic amount of stannous octoate under the protection of argon, and placing the reaction bottle in an oil bath at 110 ℃ for reaction for 48 hours. After the reaction was completed, the solvent was evaporated, dissolved in 30ml of chloroform, and precipitated with 300ml of glacial ethyl ether. The precipitate was collected, dissolved in 30ml of chloroform and precipitated with 300ml of glacial ethyl ether, which was repeated three times. The precipitate was dried in vacuo to give a white product.
3) Methyl-terminated ether polyethylene glycol-poly-L-tyrosine-polycaprolactone (CH)3Preparation of PEG-PTyr-PCL):
0.1mmol of methyl ether terminated polyethylene glycol-poly O-benzyl-L-tyrosine-polycaprolactone is dissolved in 300ml of tetrahydrofuran, 8% of palladium carbon is added, and the mixture is added into a high-pressure reaction kettle. Hydrogen gas of 5 atm was introduced, and the reaction was carried out at 40 ℃ for 72 hours. After the reaction is finished, filtering to remove palladium carbon, and performing rotary evaporation to remove tetrahydrofuran.
Claims (7)
1. A polyethylene glycol-polyamino acid-polyester triblock polymer is characterized in that the chemical structural formula of the triblock polymer is as follows:
wherein,
R1＝-CH3；
R2＝-CONHCH2CH2、-(CH2)1～5、-COCH2CH2CONHCH2CH2；
R3＝-CH2COOH、-CH2CH2COOH；
R4＝-(CH2)1～5、-CH(CH3)；
m, n and q are independently 10 to 500.
2. The process for the preparation of a triblock polymer of polyethylene glycol-polyamino acid-polyester according to claim 1, characterized in that it comprises the following steps, by weight:
(1) preparation of polyethylene glycol-polyamino acid two-block polymer
Under the protection of inert gas, weighing 0.1 molar part of dried polyethylene glycol with one end being amino, dissolving the polyethylene glycol in a reaction bottle by using 5-100 parts of anhydrous dichloromethane, weighing 0.5-10 molar parts of amino acid-N-cyclic carbonic anhydride, dissolving the amino acid-N-cyclic carbonic anhydride by using 0.5-10 parts of anhydrous N, N-dimethylformamide, then adding the mixture into the reaction bottle, reacting at the temperature of 20-50 ℃ for 12-72h, evaporating to remove the solvent, dissolving the product by using 5-30 parts of chloroform, precipitating in 50-300 parts of ethyl acetate, collecting the precipitate, dissolving in 5-30 parts of chloroform, precipitating in 50-300 parts of ethyl acetate, repeating for three times to obtain a polyethylene glycol-polyamino acid two-block polymer;
wherein, the inert gas is any one of argon, nitrogen or helium; the amino acid is any one of L-glutamic acid-gamma-benzyl ester, L-aspartic acid-beta-benzyl ester, O-benzyl-L-tyrosine or N-benzyloxycarbonyl-L-lysine;
(2) preparation of polyethylene glycol-polyamino acid-polyester triblock polymer
Weighing 0.1 molar part of polyethylene glycol-polyamino acid diblock polymer and 0.5-10 molar parts of polyester monomer in a reaction bottle, adding 5-50 parts of anhydrous dioxane, toluene or benzyl ether for dissolving under the protection of inert gas, adding 0.0001-0.1 molar part of stannous octoate, placing the reaction bottle at the temperature of 105 ℃ and 160 ℃ for reacting for 12-72h, evaporating to remove the solvent, dissolving the product with 5-30 parts of chloroform, precipitating in 50-300 parts of glacial ethyl ether, collecting the precipitate, dissolving in 5-30 parts of chloroform, precipitating in 50-300 parts of glacial ethyl ether, repeating for three times to obtain the polyethylene glycol-polyamino acid-polyester triblock polymer;
wherein, the inert gas is any one of argon, nitrogen or helium; the polyester monomer is caprolactone or lactide;
(3) polyethylene glycol-polyamino acid-polyester triblock polymer de-side group protection
Adding 0.1 molar part of polyethylene glycol-polyamino acid-polyester triblock polymer into a high-pressure reaction kettle, dissolving 100 parts of tetrahydrofuran, adding 0.8% palladium carbon, introducing hydrogen with 2-10 atmospheric pressures, reacting at the temperature of 30-50 ℃ for 24-72h, filtering to remove the palladium carbon, and performing rotary evaporation to remove the tetrahydrofuran to obtain the side group-removed polyethylene glycol-polyamino acid-polyester triblock polymer.
3. Use of a polyethylene glycol-polyamino acid-polyester triblock polymer according to claim 1, characterized in that:
(1) preparation of polyethylene glycol-polyamino acid-polyester triblock polymer nano micelle
Dissolving a polyethylene glycol-polyamino acid-polyester triblock polymer in an organic solvent, preparing a solution with the concentration of 5-20mg/ml, taking 0.5-10 parts of the solution, dripping the solution into 5-100 parts of an aqueous phase solvent at the temperature of 25-30 ℃ and the ultrasonic power of 130W and the amplitude of 65 percent, wherein the aqueous phase solvent is pure water or a phosphoric acid buffer solution with the pH value of 4-12, and shaking up; removing residual organic solvent in the aqueous phase solution by a reduced pressure rotary evaporation method to prepare the polyethylene glycol-polyamino acid-polyester triblock polymer nano micelle;
wherein the organic solvent is any one of chloroform, tetrahydrofuran or ethanol;
(2) polyethylene glycol-polyamino acid-polyester triblock polymer micelle loaded nanocrystal
a. Adding 2-10mg/ml single nano crystal dispersed in organic phase into a reagent bottle, removing organic solvent by vacuum drying or blowing dry by inert gas, and then weighing 0.001-0.5 part of nano crystal;
b. dissolving 0.005-1 part of triblock polymer in 0.5-10 parts of organic phase solvent, adding the solution into the nanocrystal, and uniformly mixing by shaking; adding the organic solution into 5-100 parts of water phase solvent at 25-30 ℃ and ultrasonic power of 130W and amplitude of 65%, wherein the water phase solvent is pure water or phosphoric acid buffer solution with pH value of 4-12, and shaking up; removing residual organic solvent in the aqueous phase solution by a reduced pressure rotary evaporation method to prepare nano micelle of the triblock polymer coated nanocrystal;
wherein, the organic phase solvent is any one of dichloromethane, trichloromethane, tetrahydrofuran or ethanol; the nanocrystal is Fe3O4Nanocrystalline, gamma-Fe2O3Nanocrystals, ZnFe2O4Nanocrystals, MnFe2O4Nanocrystals, CoFe2O4Any one of nanocrystals, FePt nanocrystals, Au nanocrystals, Ag nanocrystals, CdSe nanocrystals, CdZnS nanocrystals, CdSe/ZnS nanocrystals, or CdSe/CdS nanocrystals;
(3) polyethylene glycol-polyamino acid-polyester triblock polymer micelle loaded with hydrophobic drug
Weighing 0.001-0.5 part of hydrophobic drug and 0.005-1 part of triblock polymer, mixing and dissolving into 0.5-10 parts of organic phase solvent, uniformly mixing, adding into 5-100 parts of aqueous phase solvent at 25-30 ℃, ultrasonic power of 130W and amplitude of 65%, wherein the aqueous phase solvent is pure water or phosphoric acid buffer solution with pH value of 4-12, and shaking uniformly; removing residual organic solvent in the aqueous phase solution by a reduced pressure rotary evaporation method to prepare the nano micelle of the triblock polymer coated drug;
wherein, the organic phase solvent is any one of trichloromethane, tetrahydrofuran or ethanol; the hydrophobic drug is any one of adriamycin, paclitaxel or cisplatin;
4. the nanomicelle of polyethylene glycol-polyamino acid-polyester triblock polymer according to claim 3, characterized in that:
1) micellar internal crosslinking of polyethylene glycol-polyamino acid-polyester triblock polymer micelles
Adding 0.005-0.05 part of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride into 5 parts of polyethylene glycol-polyamino acid-polyester triblock polymer micelle solution with the concentration of 1-10mg/ml, reacting at room temperature for 10-30min, adding 0.001-0.01 part of cross-linking agent, stirring overnight, and dialyzing for 12-24h to obtain the polyethylene glycol-polyamino acid-polyester triblock polymer micelle with the cross-linked amino acid layer;
2) multifunctional nano composite particle is constructed by taking polyethylene glycol-polyamino acid-polyester triblock polymer micelle as platform
(1) The surface of micelle particles prepared by triblock polymer is modified by chemical covalent bonds to introduce micromolecular ligands, and the micelle particles are further chelated with metal ions for magnetic resonance or nuclide development;
wherein, the chemical covalent bond modification is performed by condensation reaction of carboxyl and amino, esterification reaction of carboxyl and hydroxyl, click reaction of alkynyl and azide, reaction of halide and amino, and reaction of amino and aldehyde group;
(2) introducing fluorescent molecules to the surfaces of micelle particles prepared by triblock polymers through chemical modification for fluorescent labeling;
wherein, the chemical modification is carried out by condensation reaction of carboxyl and amino, esterification reaction of carboxyl and hydroxyl, click reaction of alkynyl and azide, reaction of halide and amino, and reaction of amino and aldehyde group;
(3) the surface of micelle particles prepared by triblock polymer is introduced with functional short peptide for cell or tissue targeting through chemical modification;
wherein, the chemical modification is carried out by condensation reaction of carboxyl and amino, esterification reaction of carboxyl and hydroxyl, click reaction of alkynyl and azide, reaction of halide and amino, and reaction of amino and aldehyde group; the accessed functional short peptide is: arginine-glycine-aspartic acid or a cell-penetrating peptide.
5. The nanomicelle of polyethylene glycol-polyamino acid-polyester triblock polymer according to claim 4, characterized in that the cross-linking agent is ethylenediamine or 1, 4-butanediamine.
6. The nanomicelle of polyethylene glycol-polyamino acid-polyester triblock polymer according to claim 4, characterized in that the small molecule ligand is any one of 1,4,7, 10-tetraazacyclododecane 1,4,7, 10-tetraacetic acid, 1,4,7, 10-tetraazacyclododecane N, N', N "-triacetic acid or diethyltriaminepentaacetic acid.
7. The polyethylene glycol-polyamino acid-polyester triblock polymer nanomicelle according to claim 4, characterized in that the metal ions are: ions with +2 or +3 valence in Mn, Fe, Co, Ni, Cu, Ga, Tc and Gd.
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